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448-Temma-HeLaSPASIM/FigS11_twophoton_act_rsG_SPASIM thumbnail

448-Temma-HeLaSPASIM/FigS11_twophoton_act_rsG_SPASIM

Author reviewed
Fluorescence images of actin filaments in HeLa cell spheroids in SPA-SIM mode for two-photon activation
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Project
448-Temma-HeLaSPASIM
Dataset ID
FigS11_twophoton_act_rsG_SPASIM
Kind
Image data
Title
Fluorescence images of actin filaments in HeLa cell spheroids in SPA-SIM mode for two-photon activation
Description
Fluorescence images of actin filaments in living HeLa cell spheroids visualized with rsGamillus-S were obtained using selective-plane-activation structured illumination microscopy (SPA-SIM) with two-photon activation by a Bessel sheet. In the SPA-SIM system the aberrations make the imaging deeper parts difficult and limit the imaging depth to approximately 69 micrometre in a spheroid of ~200 micrometre in diameter. rsGamillus-S is an acid-resistance photoswitchable fluorescent protein, which owns exceptionally high on-switching rate to show the nuclei more clearly. Two-photon activation using a scanned Bessel beam produces a wider FOV and a higher axial resolution.
Folder Name
FigS11_twophoton_act_rsG_SPASIM
File Format
tif
File Size
36MB
File Separation Method
-
Figure Number
FigureS11d
Dimensions
6141x6141x1x1x1
X Scale
141.8 micrometer
Y Scale
141.8 micrometer
Z Scale
-
T Scale
-
Channel
1 channel
Quantitative Data
-
Paper(s)

Biosample

Description
HeLa cells were electroporated with rsGamillus-S plasmids to express fluorescent proteins. Actin filaments were visualized by rsGamillus-S.
Organism
Homo sapiens
(NCBITaxon:9606)
Strain
-
Cell
-
Cell line
HeLa-S3 cell (CLO:0003699)
MeSH
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Actin Cytoskeleton (D008841)
Intrinsic Variable
-
Extrinsic Variable
HeLa cells were electroporated with rsGamillus-S plasmids

Ontology

Anatomical Entity
-
UBERON
-
Biological Process
-
Cellular Component
actin filament (GO:0005884)
Molecular Function
-
MeSH Anatomy
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Actin Cytoskeleton (D008841)
MeSH BP
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Actin Cytoskeleton (D008841)
MeSH CC
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Actin Cytoskeleton (D008841)
MeSH MF
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Actin Cytoskeleton (D008841)

Genetics

Gene
-
Protein / Probe
-
Method / Genotype
electroporation

Treatments

Reagent / Compound
-
Concentration
-
Fold Dilution
-

Imaging Method

Imaging Method
structured illumination microscopy
(FBbi:00000332)
Detection Method
detection of photons
Visualization Method
rsGamillus-S
Illumination Method
light-sheet illumination
Source of Contrast
-
Imaged Parameter
-
Sample Preparation Method
-
MeSH
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Actin Cytoskeleton (D008841)

Instrument

Body
-
Module
-
Light Source
405 nm light sheet illumination
Detector
Hamamatsu Photonics ORCA-Flash 4.0 v3
Objective
Nikon CFI75 Apo 25XC W, Special Optics 54-10-7
Filter Set
-
Dichroic
Di03-R488-t3-25x36, RazorEdge LongPass488, FF01-790/SP-25